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Number of NOS-positive neurons. Examination of nitric oxide synthase- (NOS) positive cells by using NADPH-diaphorase staining compared to all plexus cells counted by nuclear fast red. Similar to the significantly reduced mRNA gene expression the number of NOS-positive neurons showed a significant decline (37.3 ± 9.1 to 20.7 ± 4.2%) in aged donors and verified the alteration of gene expression on protein level. *P ≤ 0.05.
